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Dissertations / Theses on the topic 'Leukodystrophie'
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1
Kaminski, Debora [Verfasser]. "Intrazerebroventrikuläre Enzymersatztherapie der Metachromatischen Leukodystrophie / Debora Kaminski." Bonn : Universitäts- und Landesbibliothek Bonn, 2014. http://d-nb.info/1061041875/34.
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Stein, Axel [Verfasser]. "Neuroinflammation in Mausmodellen der Metachromatischen Leukodystrophie / Axel Stein." Bonn : Universitäts- und Landesbibliothek Bonn, 2016. http://d-nb.info/1119888581/34.
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Brysch, Klaudia [Verfasser]. "De- und Remyelinisierung im Mausmodell der Metachromatischen Leukodystrophie / Klaudia Brysch." Bonn : Universitäts- und Landesbibliothek Bonn, 2019. http://d-nb.info/1200019938/34.
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Zech, Isabell [Verfasser]. "Substratreduktionstherapie der Metachromatischen Leukodystrophie: Expression der Cerebrosid-Sulfotransferase und Etablierung einer hochdurchsatzfähigen Aktivitätsbestimmung / Isabell Zech." Bonn : Universitäts- und Landesbibliothek Bonn, 2013. http://d-nb.info/1044972076/34.
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Erwes, Kim Lina [Verfasser]. "Patientenspezifische induziert pluripotente Stammzellen als Modellsystem zur Untersuchung der lysosomalen Speichererkrankung Metachromatische Leukodystrophie / Kim Lina Erwes." Bonn : Universitäts- und Landesbibliothek Bonn, 2017. http://d-nb.info/1150777737/34.
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Erwes, Kim Lina [Verfasser]. "Patientenspezifische induziert pluripotente Stammzellen als Modellsystem zur Untersuchung der lysosomalen Speichererkrankung Metachromatische Leukodystrophie / Kim Lina Erwes." Bonn : Universitäts- und Landesbibliothek Bonn, 2019. http://d-nb.info/1178162303/34.
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Weißenberg, Christine [Verfasser], and Gernot [Akademischer Betreuer] Bruchelt. "Analytik von Galaktocerebrosid-3-sulfat im Urin zur Diagnostik der Metachromatischen Leukodystrophie / Christine Weißenberg ; Betreuer: Gernot Bruchelt." Tübingen : Universitätsbibliothek Tübingen, 2011. http://d-nb.info/1161464654/34.
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Ziehme, Jakob Erdmann Sié [Verfasser], and Inge [Akademischer Betreuer] Krägeloh-Mann. "Intrafamiliäre Variabilität bei Metachromatischer Leukodystrophie – der natürliche Verlauf bei Geschwistern / Jakob Erdmann Sié Ziehme ; Betreuer: Inge Krägeloh-Mann." Tübingen : Universitätsbibliothek Tübingen, 2020. http://d-nb.info/1206933852/34.
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SCALIA, Federica. "GENETIC CHAPERONOPATHIES ASSOCIATED WITH GROUP II CHAPERONIN VARIANTS." Doctoral thesis, Università degli Studi di Palermo, 2020. http://hdl.handle.net/10447/395218.
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Genetic chaperonopathies manifest themselves from very early in life. Chaperonopathies related to neurodegenerative disorders discussed in “Introduction” section are a heterogeneous group of disorders which affect one or more of the various physiological systems, for example, the nervous system. This heterogeneity is due, in particular, to the not fully known molecular activity, which every single molecular chaperone has within a specific tissue. My general questions about them were 1) why a mutation on a molecular chaperone that is expressed by most, if not all cytotypes, seems to affect the functioning of a single physiological system? 2) why do different mutations on the same molecular chaperone cause apparently different pathologies especially in terms of clinical manifestations?
This heterogeneity limits the research approach on diseases, which now is conducted towards every single mutation without being able to generalize a unique molecular process.
I spent the first 18 months of my Ph.D. project at the SBARRO Department of Temple University in Philadelphia to study the V98I mutation on chaperone Hsp60 causing hereditary spastic paraplegia (SPG13). For a better understanding of the associated diseases, it would be highly beneficial to examine the impact of mutant chaperone genes during development, starting with fertilization and proceeding throughout the entire ontogenetic process. Zebrafish is amenable to such embryonal analysis as well as studies during adulthood. In addition, the zebrafish genome contains a wide range of genes encoding proteins similar to those that form the chaperoning system in humans. Due to the very complex roles played by Hsp60 in cell and tissue homeostasis, the gene is highly conserved during evolution. Nucleotide and amino acid sequences of Hsp60 in zebrafish have 88% of identity with its human orthologous.
The first aim of my research was the establishment of a zebrafish model as an innovative approach for the study of the molecular basis of SPG13 and define the role of missense mutations V98I.
In the last 18 months of my Ph.D. project, I was at the BIND Department of the University of Palermo and I had the possibility to study a new mutation that occurred in subunit number 5 of CCT complex. This mutation was found in a pediatric patient who is now being treated by the Department Of Sciences For The Promotion Of Health And Childhood "G. D'Alessandro" at the University of Palermo. Thus, I focused my attention on this novel variant. The main aims were 1) understanding, with the help of bioinformatics software, the type of mutation and if it causes some alteration of chaperonin molecular anatomy; 2) define the morphological changes caused by the mutation in skeletal muscle tissue.
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Stahl, Katharina. "Transgene Zebrafischmodelle zur Charakterisierung der Rolle der Astrozyten in der Entwicklung des Nervensystems und in der Leukodystrophie Alexander disease (AXD)." Diss., lmu, 2010. http://nbn-resolving.de/urn:nbn:de:bvb:19-121668.
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Strölin, Manuel [Verfasser], and Ingeborg [Akademischer Betreuer] Krägeloh-Mann. "Quantifizierung der Gehirnveränderung bei Kindern mit juveniler Metachromatischer Leukodystrophie und der Effekt der Stammzelltransplantation / Manuel Strölin ; Betreuer: Ingeborg Krägeloh-Mann." Tübingen : Universitätsbibliothek Tübingen, 2018. http://d-nb.info/1168634709/34.
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Böckenhoff, Annika [Verfasser]. "Enzymersatztherapie der metachromatischen Leukodystrophie : Untersuchungen zur Steigerung des Übertritts der Arylsulfatase A über die Blut-Hirn-Schranke durch die Verwendung von Peptidvektoren / Annika Böckenhoff." Bonn : Universitäts- und Landesbibliothek Bonn, 2013. http://d-nb.info/1044970731/34.
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Kim, Nee Na. "Therapeutic strategies in murine globoid-cell leukodystrophy (Krabbe's disease)." Thesis, University of Cambridge, 2013. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.608111.
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Piguet, Françoise. "Brain gene therapy for metachromatic leukodystrophy : towards clinical trial." Paris 5, 2011. http://www.theses.fr/2011PA05T010.
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La leucodystrophie métachromatique (LDM) est une maladie de surcharge lysosomale causée par la déficience de l’arylsulfatase A (ARSA), enzyme lysosomale impliquée dans la dégradation des sulfatides. Ce déficit enzymatique conduit à l’accumulation de sulfatides dans le système nerveux central et périphérique, entrainant une démyélinisation sévère et une dégénérescence neuronale. La forme infantile tardive est la forme la plus fréquente et la plus sévère de la pathologie, avec une évolution rapide et un décès des patients avant l’âge de 5 ans. Aucun traitement n’est actuellement disponible mais diverses stratégies thérapeutiques sont en cours d’évaluation. Du fait de la rapidité du processus neurodégénératif et de l’imperméabilité de la barrière hémato-encéphalique à l’ARSA, l’injection intracérébrale d’un vecteur viral codant l’ARSA semble l’approche thérapeutique la plus appropriée. L’injection intracérébrale d’un vecteur adeno-associé (AAV) de sérotype 5 codant l’ARSA a prouvé son efficacité pour la correction du phénotype dans le modèle murin de la pathologie. Cependant, avant d’envisager une application clinique, l’évaluation de l’efficacité et surtout de la tolérance de notre approche devait être réalisée dans un cerveau de plus grand volume. Chez le primate non-humain, nous avons démontré qu’un nombre limité d’injections intracérébrales du vecteur AAV5/ARSA entrainait une large diffusion du vecteur (37 à 46% de l’hémisphère injecté) et de l’ARSA recombinante (jusqu’à 33mm), ainsi qu’une augmentation significative de l’activité ARSA dans 50-65% de l’hémisphère injecté, sans aucun signe de toxicité. L’un des challenge majeur de la thérapie génique intracérébrale pour les maladies lysosomales est la mise en place d’une thérapie capable de traiter l’ensemble du cerveau, c'est-à-dire permettrait une large diffusion du gène et / ou de la protéine mais surtout le ciblage des cellules impliquées dans la pathologie. Le vecteur AAVrh. 10 a été démontré récemment comme permettant une meilleure diffusion de la protéine thérapeutique dans le cerveau de rongeurs après injection intracérébrale. Par conséquent nous avons décidé d’évaluer l’intérêt de ce sérotype dans notre modèle de la pathologie. L’injection de vecteur AAVrh. 10/ARSA dans le cerveau de souris LDM symptomatiques a permis une correction plus rapide (2 mois) et plus complète de la surcharge en sulfatides et des lésions histologiques en comparaison avec l’AAV5/ARSA. De plus, nous avons démontré la présence de l’enzyme thérapeutique dans les oligodendrocytes (la cellule cible de la LDM), et la quantité de C24:0-sulfatide, un isoforme de sulfatides présent exclusivement dans les oligodendrocytes, était complètement corrigée, ce qui constitue un résultat d’importance majeure d’un point de vue thérapeutique. L’utilisation d’un vecteur AAVrh. 10/GFP, nous a de plus permis de caractériser plus précisément le transport axonal et le tropisme cellulaire de l’AAVrh. 10, en démontrant notamment l’existence d’un transport rétrograde. Devant ces résultats encourageants, une étude d’efficacité et de tolérance a été réalisée chez le primate. Quatre primates ont été injecté uni ou bilatéralement avec le vecteur AAVrh. 10/ARSA de grade clinique (1. 1. 1011GC/hemisphere), dans des régions de la substance blanche déterminée au moyen d’une IRM. Dans un but clinique, la procédure chirurgicale a été optimisée afin de permettre l’injection simultanée des 12 sites, réduisant ainsi le temps de chirurgie. La tolérance de la procédure a été évalué par des tests neurologiques séquentiels et des IRM cérébrales. Les animaux ont été analysés 3 mois post injection. Nous avons démontré une diffusion importante du vecteur (jusqu’à 90% de l’hémisphère injecté) et une augmentation de l’activité ARSA allant jusqu’à 31% dans l’ensemble de l’hémisphère injecté. Comparé à nos précédents résultats avec l’AAV5, l’utilisation d’une dose 20 fois plus faible de vecteur conduit à une diffusion similaire dans l’hémisphère injecté, une quantité plus importante de vecteur dans l’hémisphère non-injecté et surtout une augmentation plus importante de l’activité ARSA. Une étude de toxicologie chez le primate est actuellement en cours avant de débuter un essai clinique de phase I/II pour des patients atteints de formes rapidement évolutives de LDM<br>Metachromatic leukodystrophy (MLD) is a lethal lysosomal storage disorder of the CNS and PNS due to a deficiency in arylsulfatase A (ARSA) enzyme that results in sulfatide accumulation, demyelination and neuronal degeneration. The late infantile form of MLD is the most frequent and severe form of the disease, death of patients occuring before the age of 5 years. No treatments are currently available for MLD, even if new therapeutic approaches are under evaluation. Given the rapid course of neurodegeneration and the impermeability of the blood brain barrier to the ARSA enzyme, direct brain delivery of ARSA through gene therapy vector seems to be the most suitable therapeutic approach. Intracerebral delivery of Adeno-Associated viral vector of serotype 5 (AAV5) encoding human ARSA was shown to efficiently correct the phenotype of the MLD mouse model. However, before moving towards clinical trial, efficiency and tolerance of the procedure needed to be evaluated in a larger size brain. In normal non-human primates, we demonstrated that a limited number of intracerebral injections of AAV5/ARSA vector led to a widespread diffusion of the vector (37-46% of the injected hemisphere) and ARSA enzyme (up to 33 mm), as well as a significant increase in ARSA activity in 50-65% of the injected hemisphere, without any signs of toxicity. One of the main challenges for brain gene therapy in LSDs is to provide a therapy allowing the treatment of the entire brain, which requires widespread delivery of the normal gene and/or protein, but also the targeting of the right type of cells. AAVrh. 10 vector was recently shown to give a better diffusion of therapeutic proteins in the rodent brain after intracerebral administration. For this reason, we decided to evaluate this serotype in our therapeutic strategy. In symptomatic MLD mice, AAVrh10/ARSA vector injection resulted in a more rapid and complete correction of sulfatide storage and brain neuropathology 2 months after intracerebral injections in comparison with AAV5/ARSA vector. Importantly, ARSA enzyme was detected in oligodendrocytes, the targeted cell in MLD, and the levels of oligodendrocyte-specific sulfatide isoforms (C24:0-sulfatide) were completely corrected, a result of major importance for therapeutic intervention. Using an AAVrh. 10/GFP vector, we also characterized more precisely the axonal transport and cell tropism of AAVrh. 10 vector and demonstrated the existence of retrograde transport of the vector. Given these encouraging results, safety and efficiency study was performed in non-human primates. GLP-grade AAVrh. 10/ARSA vector was injected in one or both hemispheres (1. 1. 1011 VG/hemisphere) in white matter areas selected using brain MRI. In a clinical perspective, surgical device was optimized to allow simultaneous infusion of 12 deposits, thus reducing time of surgical procedure. Tolerance of the procedure was assessed by sequential neurobehavioral tests and brain MRI. Animals were analyzed 3 months post-injection. We documented diffusion of the vector in up to 90% and increased activity of ARSA enzyme up to 31% in the injected hemisphere. As compared to our previous results with AAV5/ARSA, the use of a 20-fold lower dose of AAVrh. 10 vector led to increased amounts of vector in the non-injected hemisphere and increased level of ARSA activity in the injected hemisphere. Toxicological evaluation is currently ongoing in NHP, prior to a phase I/II clinical trial for rapidly evolutive forms of MLD
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Rusciano, Isabella <1990>. "Cellular signalling and morphological alterations in Autosomal Dominant Leukodystrophy." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2020. http://amsdottorato.unibo.it/9533/1/ADLD%20TESI%20dottorato%20iSABELLA%20RUSCIANO%2002.11.pdf.
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Autosomal Dominant Leukodystrophy (ADLD) is a neurodegenerative disorder that affects the central nervous system. It is a rare and late onset lethal progressive disorder without any effective treatment up to date. Genetically it is characterized by Lamin B1 gene duplication or deletion upstream the gene but the mechanisms of how the overexpression of a nuclear structural protein leads to such specific symptom as demyelination are still unclear. Considering the pivotal role that glial cells as astrocytes and oligodendrocytes and Leukemia Inhibitory Factor (LIF) have in the myelination process, this work was intended to analyse the morpho-functional aspects of different cell populations, which are engineered cellular models overexpressing Lamin B1 and primary cells deriving from ADLD patients.
The results have, to date, highlighted that the astrocytes may be pivotal in the development of the disease. Analysing the cellular ultrastructure, astrocytes overexpressing Lamin B1 show several nuclear alterations similar to the ones observed in cells deriving from ADLD patients, which, on the contrary are not present in oligodendrocytes overexpressing Lamin B1. In addiction astrocytes overexpressing Lamin B1 are not capable to produce LIF and besides the receptor (LIF-R) is downregulated, it is downregulated as well as in oligodendrocytes overexpressing Lamin B1. In both engineered models, the confirmation that this signalling pathway is altered in the cell overexpressing Lamin B1, is given to us by analysing the signalling transduction pathways downstream LIF/LIF-R. In fact, both PI3K/AKT and JAK/STAT3 axes are downregulated but surprisingly with exogenous LIF administration is possible recovered the phenotype. In particular, the toxic effect induced by Lamin B1 accumulation results in the ability of oligodendrocytes, being less affected, to completely recover the phenotype, which instead is only partial for astrocytes. Therefore, this leads us to hypothesize that astrocytes, being severely affected by Lamin B1 accumulation, lose their role of support for the oligodendrocytes in the myelination process. Finally, inflammation and ROS production was also been found in ADLD patients’ cells as alleged in literature in relation to the Lamin B1 accumulation and advancement of age. These new discoveries both as regards cellular, morphological and functional alterations, and as regards the inflammatory process and oxidative stress could pay the way for new avenues of investigation in the ADLD disease.
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Uhlenberg, Birgit [Verfasser]. "Klinische und genetische Aspekte bei Muskeldystrophie Duchenne und Becker und hypomyelinisierenden Leukodystrophien / Birgit Uhlenberg." Berlin : Medizinische Fakultät Charité - Universitätsmedizin Berlin, 2009. http://d-nb.info/1027813720/34.
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Visigalli, Ilaria. "Ex vivo gene therapy approaches for the treatment of globoid cell leukodystrophy." Thesis, Open University, 2009. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.497392.
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Globoid cell leukodystrophy (GLD) is a rare lysosomal storage disorder (LSD) due to the deficiency of the lysosomal enzyme Galactocerebrosidase (GALC). The enzymatic deficiency results in intracellular storage of undegraded metabolites in the nervous system, leading to progressive dysmyelination. We are testing the feasibility and efficacy of a gene therapy strategy based on hematopoietic stem/progenitor cells (HSPC) and lentiviral vectors (LV) in the murine model of GLD. Differently from what observed with other lysosomal enzymes, GALC gene transfer and expression in HSPC causes apoptosis and functional impairment of the transduced cells due to an inbalance of the intracellular content in bioactive sphingolipids consequent to de novo enzyme expression. Differentiated cells of the myeloid and lymphoid lineages are not affected by GALC expression, suggesting a unique sensitivity of HSPC to enzyme toxicity.
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Finnsson, Johannes. "Radiological studies of LMNB1-related autosomal dominant leukodystrophy and Marinesco-Sjögren syndrome." Doctoral thesis, Uppsala universitet, Radiologi, 2016. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-303171.
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There are approximately 6000 to 8000 rare diseases, each with a prevalence of less than 1 / 10 000, but in aggregate affecting 6 to 8% of the population. It is important to evaluate disease development and progression to know the natural course of any disease. This information can be utilized in diagnostics and in assessing effects of therapeutic interventions as they become available. This thesis describes the natural clinical history and evolution of imaging findings of two rare diseases over approximately two decades. Papers I, II and III present clinical, magnetic resonance imaging (MRI), magnetic resonance spectroscopy (MRS) and 18F-fluorodeoxyglucose positron emission tomography (FDG-PET) findings in LMNB1-related autosomal dominant leukodystrophy (ADLD). MRI was found to be very sensitive in finding pathology in patients with LMNB1-related ADLD, even before the onset of clinical symptoms. However, even patients with widespread MRI changes can have a relatively mild symptomatology and present only slight disturbances in metabolic examinations such as MRS and FDG-PET. This is compatible with relatively intact axons, even as myelin impairment is widespread. Paper IV presents clinical and MRI findings in the brain and musculature in SIL1-positive Marinesco-Sjögren syndrome (MSS), and describes a new, mild phenotype of the disease with no intellectual disabilities and only slight motor disabilities. With a 19-year-long radiological follow-up, a slow progressive atrophic process in the cerebellum and brainstem could be demonstrated. MRI of the musculature shows early involvement of the quadriceps and gastrocnemii but not the tibialis anterior, progressing to widespread atrophy in the back and upper and lower limbs at the age of 20 years. In the mildest phenotype, the most severely affected muscles were the m gluteus maximus, m sartorius, m peroneus longus, and the lateral head of the m gastrocnemius.
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Harvey, John Steven. "Metachromatic leukodystrophy : the role of non-pathogenic sequence variants in the causation of disease /." Title page, contents and abstract only, 1996. http://web4.library.adelaide.edu.au/theses/09PH/09phh341.pdf.
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Baars, Christina [Verfasser]. "Hippocampal cellular and network properties in a mouse model of metachromatic leukodystrophy / Christina Baars." Bonn : Universitäts- und Landesbibliothek Bonn, 2017. http://d-nb.info/1149154276/34.
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Schuster, Tilman [Verfasser]. "Development of Enzyme Replacement and Pharmacological Chaperone Approaches for Therapy of Metachromatic Leukodystrophy / Tilman Schuster." Bonn : Universitäts- und Landesbibliothek Bonn, 2014. http://d-nb.info/1081423668/34.
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Türkmen, Mevlude [Verfasser], Pfeifenbring [Akademischer Betreuer], Wolfgang [Gutachter] Brück, Jutta [Gutachter] Gärtner, and Thomas [Gutachter] Meyer. "Histologische Charakterisierung der Leukodystrophien und deren differentialdiagnostische Abgrenzung / Mevlude Türkmen ; Gutachter: Wolfgang Brück, Jutta Gärtner, Thomas Meyer ; Betreuer: Pfeifenbring." Göttingen : Niedersächsische Staats- und Universitätsbibliothek Göttingen, 2020. http://d-nb.info/120673163X/34.
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Pant, Devesh Chandra 1987. "Identification of the sphingolipid desaturase DEGS1 as a novel gene for a leukodystrophy in therapeutic hope." Doctoral thesis, Universitat Pompeu Fabra, 2018. http://hdl.handle.net/10803/664936.
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In spite of recent advances in understanding the genetic bases of leukodystrophies, a large number of clinical cases remain unexplained, suggesting that many leukodystrophy-associated genes have yet to be identified. Here we report 18 patients from 12 families with biallelic deleterious variants in the DEGS1 gene identified via WES. DEGS1 encodes an enzyme which catalyzes the conversion of dihydroceramide to ceramide. Common features among the patients include severe cerebellum atrophy, thinning of the corpus callosum and hypomyelination suggesting a critical role of DEGS1 in the central nervous system.
Using patient’s fibroblasts, we evidenced abnormal biochemical profiles. Knockdown of degs1 in zebrafish recapitulated the biochemical imbalance, showed impaired locomotor abilities and weak myelination. Moreover, a widely used drug for neurological disorders, fingolimod, able to normalized the toxic effects associated due to impaired DEGS1. These results pave the way to clinical translation, illustrating the transformative impact of genomics in patient care.<br>El hecho de que un gran número de casos clínicos de leucodistrofias sigan sin explicar sugiere que muchos de los genes asociados a esta enfermedad estarían aún por identificar. Secuenciando el exoma completo de 18 pacientes de 12 familias hemos encontrado variantes deletéreas bialélicas en el gen DEGS1, un gen que codifica para una enzima que cataliza la conversión de dihidroceramida a ceramida. Las características comunes entre estos pacientes incluyen atrofia grave del cerebelo, delgadez del cuerpo calloso e hipomielinización, sugiriendo un papel crítico de la proteína DEGS1 en el SNC. Sus fibroblastos envidencian un perfil bioquímico anormal.
Hemos comprobado que un modelo de knockdown del gen degs1 en pez cebra muestra capacidades locomotoras alteradas, desequilibrio bioquímico y una mielinización débil.
Hemos conseguido normalizar sus efectos tóxicos con fingolimod, un fármaco ampliamente usado para trastornos neurológicos. Son resultados que ilustran el impacto transformador de la genómica en la atención al paciente, abriendo un nuevo camino en la clínica traslacional.
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Sundblom, Jimmy. "Autosomal Dominant Leukodystrophy with Autonomic Symptoms and Rippling Muscle Disease : Translational Studies of Two Neurogenetic Diseases." Doctoral thesis, Uppsala universitet, Neurologi, 2011. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-162048.
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There is a large variety of diseases caused by single-gene mutations. Although most of these conditions are rare, together they impose a significant burden to the population. This thesis describes clinical and genetic studies of two single-gene diseases: 1) Adult-onset autosomal dominant leukodystrophy with autonomic symptoms (ADLD) caused by LMNB1 gene duplications, and characterized by autonomic, pyramidal and cerebellar symptoms. Spinal cords of patients with ADLD were studied by MRI and found to be thin, with high signal intensity in white matter. Histopathology showed loss of myelinated fibres with some reactive gliosis. DNA samples from four different families with ADLD were obtained, and the LMNB1 gene was screened for duplications. Single nucleotide polymorphism array revealed LMNB1 duplications in all ADLD families. LMNB1 mRNA and protein levels were assessed in white blood cells using quantitative polymerase chain reaction and Western blot, and increased levels of LMNB1 mRNA and lamin B1 protein could be demonstrated. We concluded that spinal cord atrophy in patients with ADLD is a valuable differential diagnostic sign, and that increased levels of LMNB1 can be detected in peripheral blood. 2) Rippling muscle disease (RMD) is caused by CAV3 gene mutations. Clinical features are percussion-induced muscle mounding, –rapid contractions and undulating muscle contractions (rippling). The CAV3 gene was sequenced in 38 members of a family with RMD. Twenty-two individuals had clinical features of RMD. No muscle weakness was seen. All patients with signs of RMD carried the p.A46T CAV3 mutation, showing that the p.A46T mutation was benign and that the diagnosis can be made clinically. In vitro contracture test results from 10 of the subjects were collected, but no association between pathological test results and RMD was found.
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Santambrogio, Sara. "Neural stem cell compartments in a mouse model of globoid cell leukodystrophy : Implication for therapeutic strategies." Thesis, Open University, 2011. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.536058.
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CASCINO, FEDERICA. "Development of chimeric lysosomal enzymes with improved bioavailability to advance gene therapy strategies for globoid cell leukodystrophy." Doctoral thesis, Università Vita-Salute San Raffaele, 2023. https://hdl.handle.net/20.500.11768/137022.
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Globoid cell leukodystrophy (GLD) is a neurodegenerative lysosomal storage disease (LSD) due to the genetic deficiency of ß-galactosylceramidase (GALC). The most frequent infantile forms display an unrelenting progression with a severe central and peripheral neurological deterioration. The only treatment option is the hematopoietic stem/progenitor cell transplant (HSPC-T), although this is poorly effective. Thus, GLD is considered an untreatable disorder.
Gene therapy (GT) provides therapeutic benefit in other LSDs that share with GLD the severe and rapid central nervous system (CNS) deterioration. However, only moderate benefit was obtained in preclinical studies for GLD and none of them was able to halt this rapid and complex multi-organ pathology. Previous studies showed that modifications to the lysosomal enzymes sequence to enhance their secretion and capability to cross the blood brain barrier (BBB) boost the GT efficacy in different animal models of LSDs. Thus, the use of a similar chimeric GALC enzyme may be crucial to successfully increase the GALC bioavailability and GT success.
We generated lentiviral vectors (LVs) encoding for the murine GALC enzyme fused with the mCherry fluorescent reporter and we engineered it by: i) replacing the GALC signal peptide (sp) with that of highly secreted lysosomal enzymes (iduronate-2-sulphatase -IDS- or -L-iduronidase -IDUA-) to increase GALC secretion; ii) adding a low-density lipoprotein receptor binding domain peptide to enhance BBB crossing. We tested generated LVs in murine GLD neural and hematopoietic stem/progenitor cells and progeny, relevant cell types in the context of GT platforms.
We showed safe overexpression and enzymatic activity of chimeric GALC enzymes in transduced GLD cells, with an advantage in terms of GALC production and secretion given by the IDSsp and even more by the IDUAsp. The chimeric enzymes were secreted by transduced cells and recaptured by GALC-deficient neural cells, mediating clearance of galactosylceramide storage in cross-corrected cells.
These results strongly support the rationale of testing the safety and efficacy of chimeric GALC enzymes in LV-mediated HSPC-GT approaches in neonatal GLD mice. To this end, we compared Busulfan and total body irradiation as myeloablative conditioning regimens in this challenging murine model and selected a protocol that has been applied in preliminary experiments to test the safety and efficacy of HSPC-GT approaches using the optimized GALC chimeric construct.
The final goal of this project is to develop novel and more effective GT strategies for this untreatable disease.<br>La leucodistrofia a cellule globoidi (GLD) è una malattia neurodegenerativa da accumulo lisosomiale (LSD) dovuta alla carenza genetica di ß-galattosilceramidasi (GALC). Le forme infantili sono le più frequenti e mostrano una progressione inarrestabile con un grave deterioramento neurologico centrale e periferico. L'unica opzione terapeutica è il trapianto di cellule staminali/progenitrici ematopoietiche (HSPC-T), sebbene sia scarsamente efficace. Pertanto, la GLD è considerata un disturbo non curabile. La terapia genica (GT) fornisce un beneficio terapeutico in altre LSD che condividono con la GLD il grave e rapido deterioramento del sistema nervoso centrale (SNC). Tuttavia, negli studi preclinici sulla GLD è stato ottenuto solo un beneficio moderato e nessuno di essi è stato in grado di arrestare questa rapida e complessa patologia multiorgano. Studi precedenti hanno dimostrato
che le modifiche alla sequenza degli enzimi lisosomiali per aumentarne la secrezione e la capacità di attraversare la barriera emato-encefalica (BBB) aumentano l'efficacia della GT in diversi modelli animali di LSD. Pertanto, l'uso di un enzima chimerico GALC simile potrebbe essere cruciale per aumentare la biodisponibilità di GALC e il successo della GT. Abbiamo generato vettori lentivirali (LV) che codificano per l'enzima GALC murino fuso con il reporter fluorescente mCherry e lo abbiamo ingegnerizzato mediante: i) sostituendo il peptide segnale (sp) di GALC con quello di enzimi lisosomiali altamente secreti (iduronato-2-solfatasi -IDS- o -L-iduronidasi -IDUA-) per aumentare la secrezione di GALC; ii) aggiungendo un peptide del dominio di legame del recettore delle lipoproteine a bassa densità per migliorare l'attraversamento della BBB. Abbiamo testato i LV in cellule staminali/progenitrici neurali ed ematopoietiche GLD murine e nella loro progenie, tipi di cellule rilevanti nel contesto delle piattaforme GT. Abbiamo dimostrato la sovraespressione sicura e l'attività enzimatica degli enzimi GALC chimerici in cellule GLD trasdotte, con un vantaggio in termini di produzione e secrezione di GALC dato dall'IDSsp e ancor più dall'IDUAsp. Gli enzimi chimerici sono stati secreti dalle cellule trasdotte e ricatturati dalle cellule neurali con deficit di GALC, mediando la degradazione degli accumuli di galattosilceramide tramite correzione incrociata. Questi risultati supportano fortemente il razionale di testare la sicurezza e l'efficacia degli enzimi GALC chimerici negli approcci HSPC-GT mediati da LV in topi GLD neonati. A tal fine, abbiamo confrontato Busulfan e l'irradiazione corporea totale come regimi di condizionamento mieloablativo in questo difficile modello murino e abbiamo selezionato un protocollo che è stato applicato in esperimenti preliminari per testare la sicurezza e l'efficacia degli approcci di HSPC-GT utilizzando il costrutto chimerico GALC ottimizzato. L'obiettivo finale di questo progetto è sviluppare strategie di GT nuove e più efficaci per questa malattia non curabile.
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Hamatani, Mio. "Altered features of monocytes in adult onset leukoencephalopathy with axonal spheroids and pigmented glia: A clue to the pathomechanism of microglial dyshomeostasis." Kyoto University, 2020. http://hdl.handle.net/2433/259005.
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Melin, Malin. "Identification of Candidate Genes in Four Human Disorders." Doctoral thesis, Uppsala : Acta Universitatis Upsaliensis, 2006. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-7344.
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SALSANO, ETTORE. "Clinical and Genetic Characterization of Leukoencephalopathies in Adults." Doctoral thesis, Università degli Studi di Milano-Bicocca, 2022. http://hdl.handle.net/10281/374739.
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Retroterra scientifico Negli adulti circa il 30-40% delle leucoencefalopatie (LKEN) (= malattie della sostanza bianca) non sono diagnosticati. I pazienti che restano non diagnosticati dopo indagini approfondite possono avare forme atipiche di malattie note, sia genetiche che acquisite, oppure nuove malattie che sono più probabilmente di origine genetica. Nel nostro lavoro abbiamo voluto esplorare l’efficienza di un approccio sistematico che include il sequenziamento del DNA con tecniche di nuova generazione (NGS) nella diagnosi di pazienti adulti con LKEN a causa ignota, e descrivere le loro caratteristiche cliniche.
Pazienti e Metodi In questo studio osservazionale analitico, abbiamo inizialmente revisionato le caratteristiche cliniche e paracliniche dei pazienti adulti (>=18 anni) con LKEN a causa ignota valutati nell’Unità di Malattie Neurodegenerative e Neurometaboliche Rare dell’Istituto Neurologico “C. Besta”, Milano, Italia, dal 2012 al 2018. Successivamente abbiamo usato un pannello genico per il sequenziamento di 142 geni associati a leucoencefalopatie ereditarie e, in un caso familiare rimasto non diagnosticato abbiamo studiato l’intero esoma.
Risultati Abbiamo identificato 57 adulti con LKEN a causa ignota (età media 43 anni, intervallo 18-72; 23 maschi, 53 con esordio tardo-adolescenziale o adulto). Trenta di loro, che abbiamo chiamato «ipomielinizzanti», presentavano un pattern di MRI suggestivo di ipomielinizzazione (lieve iperintensità in T2 e segnale T1 normale), mentre i restanti 27, che abbiamo chiamato «demielinizzanti», avevano un pattern di MRI suggestivo di demielinizzazione (marcata iperintensità in T2 e ipointensità in T1). In 13 soggetti ipomielinizzati il pannello genico ha identificato i seguenti geni: POLR3A (n = 2), POLR1C, TUBB4A, RARS1, GJA1, PLP1, GJC2, TBCD, CYP7B1, SPG11, PEX3, e PEX13, mentre in due altri pazienti lo studio dell’esoma ha portato all’identificazione di un nuovo gene malattia. Al contrario, il pannello genico ha permesso la diagnosi di un solo paziente demielinizzante (aciduria metilglutaconica associata al gene AUH). In due altri pazienti la diagnosi è stata posta o con l’analisi diretta di un singolo gene-malattia dopo revisione dei dati clinico-neuroradiologici (leucodistrofia metacromatica associata al gene PSAP) o con una biopsia di cute dopo la negatività del pannello genico (malattia da inclusi intranucleari neuronali, NIID). Tre pazienti (uno ipomielinizzante, due demielinizzanti) avevano malattie acquisite che mimavano una leucodistrofia: una vasculite primaria diagnostica con la biopsia cerebrale senza analisi genetiche e rare varianti di sclerosi multipla, diagnosticate dopo la negatività del pannello. Infine, in otto pazienti con una LKEN scoperta incidentalmente che sono rimasti senza manifestazioni cliniche per anni, il pannello genico ha dato esito negativo.
Conclusioni Negli adulti un pattern ipomielinizzante caratterizza un gran numero (circa il 50%) di LKEN a causa ignota. Le LKEN ipomielinizzanti ad esordio tardivo sono molto più comunemente dovute a geni associati alle forme precoci, come POLR3A e TUBB4A, o possono essere dovute a geni associati a paraplegie spastiche ereditarie, come CYP7B1 e SPG11, a geni associati a disordini perossisomiali, come PEX3 e PEX13, o anche a nuovi geni malattia. Fra le forme demielinizzanti solo molto poche sono diagnosticate con un pannello genico se i dati clinico-paraclinici non orientano verso una diagnosi specifica. Occasionalmente, varianti atipiche di malattie acquisite della sostanza bianca possono mimare una LKEN genetica con caratteristiche demielinizzanti o ipomielinizzanti alla risonanza. Infine, un sottogruppo di LKEN demielinizzanti caratterizzato dalla mancanza di manifestazioni cliniche e dall’assenza di mutazioni in geni associati a forme note di LKEN può costituire una nuova entità che abbiamo chiamato leucoencefalopatia diffusa subclinica.<br>Background In adults, many cases (about 30-40%) of leukoencephalopathies (LKENs), i.e. white matter (WM) diseases, are without definitive diagnosis. Patients who remain undiagnosed despite extensive investigations may have atypical forms of known acquired or genetic diseases, or novel diseases more likely genetic in nature. Aims of our work were to explore the efficiency of a systematic approach, including next generation sequencing (NGS), in the diagnosis of a cohort of adult patients with LKEN of unknown cause, and to describe their clinical features.
Patients and Methods In this analytical observation study, we first reviewed the clinical and laboratory features of the adult patients (age >= 18 years) with undiagnosed LKEN assessed at the Unit of Rare Neurodegenerative and Neurometabolic Diseases of the Istituto Neurologico “C. Besta”, Milan, Italy, from 2012 to 2018. A targeted-gene panel sequencing (TGPS) was subsequently used to investigate 142 genes responsible for genetic LKENs, and a whole-exome sequencing (WES) was performed in one familial case remained undiagnosed.
Results We identified 57 adult patients with LKEN of unknown cause (mean age 43 years, range 18-72; 23 males; 53 with late-adolescence or adult-onset). Thirty of them, henceforward called hypomyelinating leukoencephalopathies (HypoLKENs), presented an MRI pattern suggestive of hypomyelination (mild T2-hyperintensity and normal T1 signal), whereas the remaining 27 (henceforward called demyelinating leukoencephalopathies, DemLKENs) had an MRI pattern suggestive of demyelination (prominent T2-hyperintensity and prominent T1-hypointensity). In 13 HypoLKENs, TGPS identified the disease-causing genes, i.e., POLR3A (n = 2), POLR1C, TUBB4A, RARS1, GJA1, PLP1, GJC2, TBCD, CYP7B1, SPG11, PEX3, and PEX13, while in two further patients, WES led to the identification of a novel disease-causing gene (preliminarily called GENE_A). In contrast, TGPS identified the disease-causing gene (i.e., AUH) in only one (out of 27) DemLKEN patient affected by methylglutaconic aciduria type 1. In two other DemLKEN patients, the diagnosis was made on the basis of their clinical and MRI features directly by single gene analysis (PSAP-related metachromatic leukodystrophy), or by skin biopsy after negative results of TGPS (neuronal intranuclear inclusion disease, NIID). Three patients (one with HypoLKEN and two with DemLKEN) had acquired diseases mimicking a leukodystrophy, i.e., a primary cerebral vasculitis (diagnosed by brain biopsy without genetic analyses) and rare variants of multiple sclerosis, diagnosed after negative results of TGPS. Finally, in eight subjects with an incidentally found DemLKEN who remained without clinical manifestations over a long period of time, no mutation was found by TGPS.
Conclusions In adults, a hypomyelinating pattern characterizes a large number (about 50%) of LKENs of unknown cause. HypoLKENs are most commonly due to genes causing severe early-onset hypomyelinating leukodystrophies (HLDs), such as POLR3A and TUBB4A, or can be due to genes associated with hereditary spastic paraplegias, such as CYP7B1 and SPG11, peroxisomal biogenesis disorders, such as PEX3 and PEX13, or even novel disease-causing genes. Among the DemLKENs of unknown cause, only very few are diagnosed by TGPS if clinical and paraclinical data pointing toward specific diagnoses are lacking. Occasionally, atypical variants of acquired WM diseases can mimic a genetic leukoencephalopathy with demyelinating or hypomyelinating features on MRI. Finally, a subset of DemLKENs characterized by lack of neurological manifestations and no mutation after comprehensive NGS testing may constitute a novel entity we termed subclinical diffuse leukoencephalopathy (SDL).
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SFERRA, ANTONELLA. "RNF220: a novel ubiquitin E3 ligase associated to autosomal recessive leukodystrophy with ataxia and deafness AR-LAD: is involved in nuclear lamina integrity control." Doctoral thesis, Università degli Studi di Roma "Tor Vergata", 2012. http://hdl.handle.net/2108/202231.
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Autosomal recessive leukodystrophy with ataxia and deafness (AR-LAD) is a rare white matter disease that slowly develops cerebellar ataxia and deafness. As disease progresses, the patients develop a fatal cardiomyophathy that strongly recalls that observed in patients affected by laminopathies, a group of disorders caused by mutations in A-type lamins (Lamin A/C), the major components of nuclear lamina. Linkage analysis and positional cloning experiments allowed us to identify the causative gene as RNF220 and two different homozygous missense mutations in highly conserved residues in exon 8 (c.1088G>A and c.1094G>A) (Dr. Bertini, unpublished data). RNF220 gene encodes for a RING finger ubiquitin ligase with a predicted molecular weight of 62.7 kDa highly expressed in liver, kidney and brain. Ubiquitin ligases catalyze the covalent attachment of ubiquitin to the substrate and play a key role in the ubiquitin-mediated proteolysis, however to date the role of RNF220 and the effects of pathological mutations associated with AR-LAD still need to be demonstrated. Our study has been focused on the functional characterization of RNF220, particularly we proposed to: 1) study the expression and subcellular distribution of RNF220, 2) verify if RNF220 may be involved in protein degradation via UPS and eventually determinate the spatial relationship between RNF220 and the different compartments associated with the proteasome, 3) investigate the involvement of RNF220 in the maintenance of nuclear lamina integrity, 4) investigate the possible interaction of RNF220 with molecules involved in the autophagic pathway. The results of this study have shown that: 1) RNF220 is an ubiquitin ligase with a nuclear distribution, 2) RNF220 co-localized with the 20S subunit, the catalytic core of the 26S proteasome, in nuclear speckles where is likely involved in protein degradation via the ubiquitin system, 3) RNF220 is a key player in the maintenance of nuclear integrity as shown by alteration/delocalization of Lamin A/C in AR-LAD fibroblast and in COS-1 cells ectopically expressing RNF220, 3) RNF220 forms a complex with Ambra1, a novel autophagic molecule, and co-localized with it in the nucleus, suggesting a RNF220 implication in autophagic pathway. Taken together, our results suggest that RNF220, the defective protein of AR-LAD is a key player of a novel degradation complex at nuclear level, connecting autophagic and proteosomal pathways and playing a crucial role in maintaining nuclear integrity.
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López, Hernández Tania. "Aproximaciones bioquímicas y celulares a la fisiopatología de la Leucoencefalopatía Megalencefálica." Doctoral thesis, Universitat de Barcelona, 2012. http://hdl.handle.net/10803/83531.
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La Leucoencefalopatía Megalencefálica con Quistes Subcorticales (MLC) es un tipo raro de leucodistrofia vacuolizante, que presenta como principales características clínicas macrocefalia, deterioro de las funciones motoras, epilepsia y retraso mental medio. Sin embargo, el diagnóstico de MLC se confirma mediante imágenes de resonancia magnética, donde el encéfalo se presenta atrofiado e hinchado, muestra una sustancia blanca anormalmente difusa y hay presencia de quistes subcorticales. Desde el punto de vista fisiopatológico, una biopsia obtenida de un paciente de MLC muestra la presencia de numerosas vacuolas situadas en las láminas más externas de la mielina.
Se ha encontrado un primer gen responsable de la enfermedad en el 75% de los pacientes afectados, denominado MLC1. Se han descrito alrededor de 60 mutaciones, aunque existen pacientes que manifiestan las características clínicas de la enfermedad pero no presentan mutaciones en MLC1 ni presentan ligamiento con su locus, sugiriendo que existe al menos otro gen involucrado en la enfermedad. En el 25% de pacientes restantes, la enfermedad se manifiesta de dos maneras diferentes: en un caso, los enfermos presentan las mismas características clínicas que los pacientes con mutaciones en MLC1; y en el otro, presentan síntomas transitorios y los pacientes mejoran, llegando incluso a que la enfermedad remitiera.
El gen MLC1 codifica para una proteína transmembrana que lleva el mismo nombre. Su función es todavía desconocida. Aunque muestra un bajo grado de homología con el canal de potasio Kv1.1 no se ha podido detectar actividad de canal iónico en diferentes sistemas heterólogos. No obstante, dicha homología, su confinamiento en la membrana plasmática y el fenotipo característico vacuolizante de los pacientes sugieren que la proteína podría estar mediando la translocación de iones a través de la superficie celular. El total desconocimiento del rol preciso de la proteína MLC1 ha imposibilitado el entendimiento del mecanismo patofisiológico de la enfermedad, y por ello, no se ha podido desarrollar ningún tratamiento efectivo para los pacientes afectados.
Es por ello que nuestro grupo quiso apostar por estrategias innovadoras (combinación de bioquímica y genética) para poder encontrar otros genes responsables de la enfermedad. Usando técnicas de purificación por afinidad combinada con métodos de proteómica cuantitativa encontramos a GlialCAM como una proteína que estaba asociada con MLC1. Es por eso que decidimos estudiar (en colaboración) si los pacientes que no tenían mutaciones en MLC1 podían presentar mutaciones en GLIALCAM. Tras el análisis de 40 de estos pacientes encontramos que cuando los enfermos tenían las características clínicas típicas de MLC presentaban dos mutaciones en GLIALCAM (herencia recesiva); mientras que en el caso de aquellos que mejoraban a lo largo del tiempo, éstos solo presentaban una mutación (herencia dominante), demostrando que GLIALCAM es el segundo gen de MLC. En este estudio también se ha podido determinar que mutaciones dominantes en GLIALCAM podían también causar otras enfermedades como la macrocefalia familiar benigna y la macrocefalia con retraso mental, con o sin autismo.
Estudios bioquímicos posteriores han permitido avanzar en el entendimiento de la relación que existe entre MLC1 y GlialCAM. Así se ha demostrado que GlialCAM actúa como una molécula escolta, necesaria para localizar específicamente a MLC1 en uniones celulares. De esta forma pudimos descubrir que las mutaciones en GLIALCAM provocaban un defecto en el tráfico de la proteína debido a una deficiente oligomerización. Como consecuencia, estas mutaciones provocaban la deslocalización de los complejos de MLC1-GlialCAM en las uniones astrocitarias. De forma interesante, GlialCAM permite estabilizar la proteína MLC1, sugiriendo nuevas aproximaciones terapéuticas para los pacientes afectos con MLC.
Tras el descubrimiento de GlialCAM como segundo gen de MLC gracias a la aproximación proteómica, y tras comprobar que no todo GlialCAM estaba asociado a MLC1, nos planteamos volver a realizar estudios de proteómica para intentar encontrar posibles proteínas que pudiesen estar interaccionando con GlialCAM. De esta manera encontramos que el canal de cloruro ClC-2, estaba asociado con GlialCAM, y pudimos comprobar que GlialCAM también actuaba como molécula escolta para localizar específicamente a ClC-2 en las uniones entre células. Además, también era capaz de modificar sus propiedades de canal, así como aumentar su función, demostrándose interacción directa entre ambas proteínas. Igualmente que para el caso de MLC1, las mutaciones encontradas en GLIALCAM fallaban en la capacidad de concentrar a ClC-2 en las uniones astrocitarias. Por tanto, la función de GlialCAM podría ser necesaria para agrupar tanto a MLC1 como a ClC-2 en tales uniones, particularmente en los pies terminales astrocitarios, donde podrían estar llevando a cabo su función. ClC-2 podría ser necesario para desarrollar un flujo de Cl- transcelular o para compensar gradientes electroquímicos iónicos que pueden estar ocurriendo en dichas uniones durante cambios en la osmolaridad. El descubrimiento de GlialCAM como una subunidad auxiliar de ClC-2 incrementa la compleja regulación de este canal y proporciona nuevas ideas acerca del papel que ClC-2 puede estar desempeñando en las células gliales así como se sugiere que pueda estar involucrado en la fisiopatología de MLC.<br>Megalencephalic leukoencephalopathy with subcortical cysts (MLC) is a leukodystrophy characterized by early-onset macrocephaly and delayed-onset neurological deterioration. Recessive MLC1 mutations are observed in 75% of patients with MLC. Genetic-linkage studies failed to identify another gene. We have showed that some patients without MLC1 mutations display the classical phenotype; others improve or become normal but retain macrocephaly. To find another MLC-related gene, we used quantitative proteomic analysis of affinity-purified MLC1 as an alternative approach and found that GlialCAM, an IgG-like cell adhesion molecule, is a direct MLC1-binding partner. Analysis of 40 MLC patients without MLC1 mutations revealed multiple different GLIALCAM mutations. Patients with the classical phenotype had two mutations, and patients with the improving phenotype had one mutation. In addition, patients with dominant GLIALCAM mutations, could also had macrocephaly and mental retardation with or without autism. Therefore, we found that GLIALCAM is the second gene found to be mutated in MLC.
Furthermore, we demonstrated that GlialCAM functions as an MLC1 beta-subunit, needed for proper localization of MLC1 in cell-cell junctions. We also demonstrated that MLC1 and GlialCAM form homo- and hetero-complexes and that MLC-causing mutations in GLIALCAM mainly reduce the formation of GlialCAM homo-complexes, leading to a defect in the trafficking of GlialCAM alone to cell junctions. GLIALCAM mutations also affect the trafficking of its associated molecule MLC1, explaining why GLIALCAM and MLC1 mutations lead to the same disease: MLC.
In this thesis, we also identify GlialCAM as a chloride channel ClC-2 binding partner. GlialCAM and ClC-2 colocalize in Bergmann glia, in astrocyteastrocyte junctions at astrocytic end-feet around blood vessels, and in myelinated fiber tracts. GlialCAM targets ClC-2 to cell junctions, increases ClC- 2 mediated currents, and changes its functional properties. Disease-causing GLIALCAM mutations abolish the targeting of the channel to cell junctions. Hence, we describes the first auxiliary subunit of ClC-2 and suggests that ClC-2 may play a role in the pathology of MLC disease.
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Arnedo, Llena Tanit. "Paper del canal de clorur CIC-2 en les patologies de la mielina." Doctoral thesis, Universitat de Barcelona, 2015. http://hdl.handle.net/10803/310220.
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La Leucoencefalopatia Megalencefàlica amb Quists subcorticals (MLC) és un tipus rar de leucodistròfia vacuolitzant de progressió lenta, que presenta com a principals característiques clíniques macrocefàlia acusada durant els primers anys de vida, deteriorament de les funcions motores, epilèpsia i retard mental de grau mig. Actualment encara es desconeix el mecanisme fisiopatològic de la malaltia, i per tant ni hi ha cap tractament possible per als pacients. S’han descrit dos gens implicats en la malaltia MLC. El primer gen descobert s’anomena MLC1 i codifica per una proteïna de membrana que porta el mateix nom. El segon gen s’anomena GLIALCAM i codifica per una proteïna transmembrana de tipus I que també porta el mateix nom. S’ha decrit que la proteïna GlialCAM actua com a subunitat ß de MLC1 ja que es capaç de dirigir-la i concentrar-la a les unions cel·lulars. Per altra banda, GlialCAM també s’ha descrit com a subunitat auxiliar del canal de Cl- ClC-2 ja que és capaç de modificar les propietats d’activació i rectificació del canal. Recentment s’ha descrit mutacions en ClC-2 associades a un tipus de leucodistròfia vacuolitzant. L’objectiu general d’aquesta Tesi és avançar en la comprensió del possible mecanisme d’acció i la funció de la proteïna GlialCAM i ClC-2 i així aprofundir en el coneixement de en el seu paper en les cèl·lules glials així com en la fisiopatologia de les leucoencefalopaties vacuolitzants. Per a realizar aquest objectiu es van realitzar estudis d’estructura-funció de GlialCAM mitjançant la caracterització bioquímica i funcional de noves mutacions en GLIALCAM associades a MLC. S’ha obtingut una classificació de les mutacions en GLIALCAM en funció del defecte que presentaven. S’ha descrit mutacions defectives en la expressió proteica, defectives en la homooligomerització i en el tràfic a les unions cel·lulars, defectives únicament en el tràfic cel·lular, mutacions sensibles a la manca de MLC1 i mutacions defectives en la internalització de la proteïna. Paral·lelament es va aprofundir en la relació bioquímica entre GlialCAM i ClC-2 a partir de l’estudi bioquímic i funcional de mutacions en CLCN2 associades a leucoencefalopaties vacuolitzants. S’ha observat que GlialCAM augmenta la funció del canal ClC-2 a través de la modificació del gating del canal i de la estabilització de ClC-2 a la membrana plasmàtica però no sembla que millori la sortida de ClC-2 del reticle endoplasmàtic. A més, aquesta estabilització requereix de la formació d’homocomplexes de GlialCAM. Per últim, es va generar i caracteritzar un model knock-down de la proteïna ClC-2 per aprofundir el paper de ClC-2 en els astròcits. Així com també es va avançar en la relació bioquímica i funcional entre GlialCAM i ClC-2 en la fisiologia astrocitària. S’ha descrit que en astròcits en cultiu en condicions d’alta concentració de K+, similar al que succeiria en situacions d’alta activitat neuronal, ClC-2 es transloca de l’aparell de Golgi a les membranes cel·lulars, modificant les seves propietats funcionals per l’efecte de GlialCAM. En canvi, en astròcits deficients de MLC1, ClC-2 es troba retinguda citoplasmàticament. Aquest fet, indicaria que aquestes proteïnes podrien tenir un paper en el procés de sifoneig del K+, i per tant, la deslocalització de ClC-2 podria donar lloc a un desordre en l’homeòstasi d’aigua i ions.<br>Megalencephalic Leukoencephalopathy with subcortical Cysts (MLC) is a rare type of vacuolating leukodystrophy. Currently still unknown pathophysiological mechanism of the disease, and therefore there is no effective treatment possible for patients. There are two genes involved in the MLC disease. Gene was first discovered was MLC1 and this encodes for a membrane protein with the same name. The second gene is called GLIALCAM and encodes for a transmembrane protein type I that also carries the same name. In our group is has been described that GlialCAM acts as a protein ß subunit of MLC1 because it is able to direct and concentrate in the cellular junctions. Moreover, GlialCAM also act as auxiliary subunit of CLC-2 Cl channel as it is capable of modifying the activation and rectification properties of the channel. Recently, mutations in ClC-2 have been associated with a rare type of vacuolating leukodystrophy. The principal aim of this study is to advance in the knowledge of GlialCAM and ClC-2 in the glial cells and into the pathogenesis of vacuolating leukodistrophies. To accomplish the study, the group performed a biochemical and funcional characterization of new mutations in GLIALCAM associated with MLC. We suggest that the HEPACAM mutations described up to now can be classified in several groups. Some mutations can affect GlialCAM protein expression, affect its ability to cis-homooligomerize and consequently reduce their localization in cell–cell junctions. Some mutations can affect specifically only transinteractions between GlialCAM molecules or may be unstable without MLC1 and finally some mutations affect the protein internalization. Parallel progress was made in the biochemical relationship between GlialCAM and ClC-2 from biochemical and functional studies of mutations in CLCN2 associated with vacuolating leukodystrophies. GlialCAM has been observed to increase ClC-2 function by the modification of its gating and the stabilization of ClC-2 in the plasma membrane. In addition, the stabilization requires a previous formation of GlialCAM’s homocomplexes. Finally, a knock-down model of the CLC-2 protein was generated and characterized to deepen the role of CLC -2 astrocytes. As well as progress was made in biochemistry and functional relationship between CLC -2 and GlialCAM in the astrocitic physiology. It has been reported that astrocytes cultured in conditions of high concentrations of K + , similar to what happens in situations of high neuronal activity , CLC -2 translocates from the Golgi apparatus to the cell membrane , changing the its functional properties for the purpose of GlialCAM . However, in MLC1 deficient astrocytes, CLC -2 is retained intracellularly. This would indicate that these proteins could play a role in the potassium siphoning, and therefore the relocation of CLC -2 could lead to disorder in the homeostasis of water and ions.
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Elitt, Matthew S. "DISEASE MODELING AND THERAPEUTIC DEVELOPMENT FOR PELIZAEUS-MERZBACHER DISEASE." Case Western Reserve University School of Graduate Studies / OhioLINK, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=case1536687505814955.
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Depiets, Bérengère. "Etude physiopathologique de modèles murins de leucodystrophies dysmyélinisantes et approche thérapeutique." Thesis, Clermont-Ferrand 1, 2012. http://www.theses.fr/2012CLF1MM04/document.
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Les mutations du gène des protéolipoprotéines, PLP1, codant des protéines structurales majeures de la myéline du système nerveux central : PLP et DM20, sont responsables d'un sous-groupe de leucodystrophies dysmyélinisantes liées à l'X. La forme la plus sévère, la maladie de Pelizaeus-Merzbacher (PMD), induite principalement par des duplications du gène conduit à une hypomyélinisation majeure ; tandis que la forme la plus modérée, la paraplégie spastique de type 2 (SPG2), induite par des mutations non-sens ou des délétions du gène conduit à une myéline mal compactée et une dégénérescence axonale tardive. Ce travail de thèse porte sur la caractérisation phénotypique de souris transgéniques mâles présentant une invalidation du gène Plp1 (souris Plp null), ainsi que des femelles hétérozygotes pour cette mutation et surexprimant le gène Plp1 (souris PLOA), modèles des mères transmettrices de ces maladies. Une étude longitudinale du comportement de ces souris a été réalisée et a permis de mettre en évidence chez les souris mâles Plp null l'apparition de troubles moteurs, sensitifs et cognitifs, qui ont pu ensuite être reliés à des anomalies d'expression (1) de marqueurs astrocytaires ou microgliaux et de neuropeptides impliqués dans la douleur au niveau de la moelle épinière, (2) de marqueurs ayant un rôle dans les processus cognitifs dans le cerveau et notamment dans certaines régions de l'hippocampe et (3) à des altérations des vitesses de conduction nerveuse. Chez les femelles mutées Plp1, les anomalies comportementales semblent liées au génotype, avec le développement de symptômes uniquement chez les mères porteuses de mutation modérée. Depuis quelques années, un ensemble de données évoquent un rôle de la substance blanche, et notamment la myéline, dans les fonctions comportementales et cognitives. Les résultats obtenus au cours de ce travail confirment l'intérêt des souris Plp null pour mieux comprendre ce rôle. De plus, les nombreuses similarités identifiées entre les modèles animaux et la pathologie humaine permettent d'envisager l'utilisation de ces modèles pour l'évaluation de nouvelles stratégies thérapeutiques. Nous avons ainsi évalué l'efficacité d'un neuroleptique atypique sur les troubles comportementaux des souris mâles Plp null<br>Mutations of the proteolipoprotein gene, PLP1, coding the major structural proteins of the central nervous system, PLP and DM20, are responsible of some X-linked dysmyelinating leukodystrophies. The most severe form, the Pelizaeus-Merzbacher disease (PMD), due to gene duplications, causes a major hypomyelination ; while the moderate form, the spastic paraplegia type 2 (SPG2), due to non-sense mutations or gene deletions, leads leading to unpacked myelin and late axonal degeneration. This thesis work focuses on phenotypic characterization of transgenic male mice with Plp1 invalidation (Plp null mice), together with heterozygous females for this mutation and overexpressing Plp1 (PLOA mice), models of carrier mothers of these diseases. A longitudinal study on mice behavior was performed and allowed to highlight in Plp null male mice, the onset of motor, sensitive and cognitive defects, then linked to expression abnormalities of (1) astrocytic or microglial markers and neuropeptides involved in painful processes in spinal dorsal horn, (2) markers implied in cognitive processes in brain and especially some hippocampus regions, (3) alterations of nerve conduction velocities. In Plp1-mutated females, behavior abnormalities seem to be related to genotype, with development of symptoms only in females carrying moderate mutation. Since few years, data suggest a role of white matter, and particularly myelin, in cognitive and behavioral functions. Results of this study confirm the interest of Plp null mice to better understand this role. Further, similarities identied between animal models and human pathology, allow to consider these models to assess new therapeutic perspectives. We thus assessed the efficiency of a typical neuroleptic on Plp null mice behavioral alterations
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Redin, Claire. "NGS-based approaches for the diagnosis of intellectual disability and other genetically heterogeneous developmental disorders." Thesis, Strasbourg, 2014. http://www.theses.fr/2014STRAJ129/document.
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Certaines maladies héréditaires monogéniques sont caractérisées par une grande hétérogénéité génétique. Chez des individus présentant un phénotype clinique similaire, les mutations causales peuvent être retrouvées dans un des gènes parmi un sous-ensemble décrits comme impliqués dans la maladie. Cette hétérogénéité génétique limite considérablement les offres diagnostiques pour les patients, et une majorité reste sans diagnostic moléculaire. Nous avons développé une approche diagnostique alternative par séquençage à haut débit ciblé (ciblant spécifiquement les régions codantes des gènes d’intérêt par capture d’exons), au travers de trois pathologies génétiquement hétérogènes : le syndrome de Bardet-Biedl (19 gènes décrits), les leucodystrophies (50 gènes), et la déficience intellectuelle (>400 gènes). Au vu de son efficacité dans le syndrome de Bardet-Biedl et la déficience intellectuelle (80% et 25% de mutations détectées respectivement, soit des taux nettement supérieurs à ceux des méthodes précédentes), elle est depuis appliquée en routine diagnostique. Au-delà du diagnostic, cette approche permet de manière non biaisée de revoir la contribution de chacun des gènes dans la pathologie et donc d’identifier les gènes récurrents, et d’établir de nouvelles corrélations génotype/phénotype<br>Some monogenic disorders are characterized by a vast genetic heterogeneity. In individuals with similar clinical phenotype, causative mutations can be found in one gene from a subset described as implicated in the disease. Such genetic heterogeneity limits considerably the diagnostic offer for the patients, and a majority is left without molecular diagnosis. We developed an alternative diagnostic approach by targeted high throughput sequencing (specific to the coding regions of genes of interest by a technique of exon capture) through three genetically heterogeneous disorders: Bardet-Biedl syndrome (19 genes reported), leukodystrophies (50 genes), and intellectual disability (>400 genes). In light of its efficiency, this approach has since been implemented in diagnostic routine for Bardet-Biedl syndrome and intellectual disability (80% and 25% of diagnostic yields respectively, significantly higher than those of previous methods). Beyond diagnosis, this approach allows unbiased means to assess the contribution of each gene in the disease and highlight recurrent genes, and establish new correlations genotype to phenotype, overall providing much insight in the genetics of a particular disease
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Avola, Rosanna. "Dynamic expression of aquaporins in physiological and pathophysiological in vitro models." Doctoral thesis, Università di Catania, 2017. http://hdl.handle.net/10761/3620.
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Water is the main component of biological fluids and a prerequisite of all organisms living. In 1987, Agre isolated a new integral membrane protein acting as a channel that mediates the water flux and uncharged solutes across biological membranes. This protein was called aquaporin1 and ever since its discovery, more than 300 homologues have been identified in animal, bacteria and plant. In human have been discovered 13 aquaporins (AQPs) isoform (AQP0-AQP12) widely distributed in various epithelia and endothelia where are important actors of fluid homeostasis in secretory and absorptive processes in response to an osmotic or pressure gradient. In the human brain nine aquaporin subtypes (AQP1, 2, 3, 4, 5, 7, 8, 9, and 11) have been recognized and partially characterized, but only three aquaporins (AQP1, 4, and 9) have been clearly identified in vivo. This discovery highlighted the concept of the important role of AQPs in all brain functions and of the dynamics of water molecules in the cerebral cortex. Additionally, AQPs relieved an important role in glial control and neuronal excitability, such as in brain structure and general development. However, a clearer understanding of specific function and distribution of water channels in adult or in development brain requires a more detailed elucidation. Some of these findings are limited from the complexity of direct investigation, inaccessibility of the neural tissue, and hence difficulty in obtaining a brain biopsy, until after the death of an individual. In this sense, several past and present in vitro models have been used to provide important clues about many processes, such as brain development, neurotoxicity, inflammation, pathogenic mechanisms of the diseases and potential pharmacological targets. In the Chapter I, we have reviewed some in vitro approaches used to investigate the mechanisms involved in Krabbe disease with particular regard to the cellular systems employed to study processes of inflammation, apoptosis and angiogenesis. In this study, we used some in vitro methods with the aim to update the knowledge on stem cells biology and to provide a relationship between aquaporins expression and cellular differentiation. In particular, we have analysed the differentiation of human mesenchymal stem cells from adipose tissue (AT-MSCs) into neural phenotypes and SH-SY5Y neuroblastoma cell line into physiological and pathophysiological dopaminergic neurons. In the Chapter II, we have reported the results of the expression of AQP1, 4, 7, 8 and 9 at 0, 14, and 28 days in AT-MSCs during the neural differentiation by immunocytochemistry, RT-PCR and Western blot analysis. Our studies demonstrated that AT-MSCs could be differentiated into neurons, astrocytes and oligodendrocytes, showing reactivity not only for the typical neural markers, but also for specific AQPs in dependence from differentiated cell type. Our data revealed that at 28 days AT-MSCs express AQP1, astrocytes AQP1, 4 and 7, oligodendrocytes AQP1, 4 and 8, and finally neurons AQP1 and 7. In the Chapter III, we have examined the possible involvement of AQPs in a Parkinson s disease-like cell model. For this purpose, we used SH-SY5Y cell line, differentiated in dopaminergic neurons with retinoic acid (RA) and phorbol 12-myristate 13-acetate (MPA) alone or in association. The vulnerability to dopaminergic neurotoxin 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) and H2O2 was evaluated and compared in all cell groups. We found that the vulnerability of cells was linked to dynamic changes of AQP4 and AQP9. The data described here provides fundamental insights on the biology of the human mesenchymal stem cells and significant evidences on the involvement of AQPs in a variety of physiological and pathophysiological processes. This suggests their possible application as markers, which may be helpful in diagnosing as well as in the understanding of neurodegenerative diseases for future therapeutic approaches.
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Weiler, Nina [Verfasser]. "Entwicklung eines Zellkulturmodells zur metachromatischen Leukodystrophie durch Transduktion glialer Zellen mit lentiviralen Vektoren / Nina Weiler." 2007. http://d-nb.info/984625321/34.
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Graf, Franziska. "Morphometrische Analyse des Axonwachstums bei Arylsulfatase A- und Ceramidgalaktosyltransferase-defizienten Mäusen als Modell der humanen metachromatischen Leukodystrophie." 2005. http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&doc_number=015430894&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA.
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Kassmann, Celia Michèle [Verfasser]. "Konditionale Inaktivierung von Peroxisomen im Nervensystem der Maus : ein transgenes Modell einer humanen Leukodystrophie / vorgelegt von Celia Michèle Kassmann." 2006. http://d-nb.info/979178746/34.
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Stahl, Katharina [Verfasser]. "Transgene Zebrafischmodelle zur Charakterisierung der Rolle der Astrozyten in der Entwicklung des Nervensystems und in der Leukodystrophie Alexander disease (AXD) / vorgelegt von Katharina Stahl." 2010. http://d-nb.info/1008344087/34.
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Matthes, Frank [Verfasser]. "Enzymersatztherapie der metachromatischen Leukodystrophie : Untersuchungen zur therapeutischen Wirksamkeit rekombinanter humaner Arylsulfatase A und zum transendothelialen Übertritt über die Blut-Hirn-Schranke / vorgelegt von Frank Matthes." 2010. http://d-nb.info/1009935798/34.
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Lin, Dar-Shong, and 林達雄. "CNS-targeted Gene Therapy in Globoid Cell Leukodystrophy." Thesis, 2013. http://ndltd.ncl.edu.tw/handle/k5zrm2.
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博士<br>國立臺北科技大學<br>工程科技研究所<br>101<br>Globoid cell leukodystrophy (GLD) is a devastating lysosomal storage disease caused by deficiency of the enzyme galactocerebrosidase (GALC). Currently, there is no definite cure for the GLD. Several attempts with CNS-directed gene therapy in twitcher mice (a murine model of GLD) demonstrated restricted expression of GALC activity in CNS and failure of therapeutic efficacy in cerebellum and spinal cord, resulting in various degrees of correction of biochemical, pathological and clinical phenotype. More recently, twitcher mice receiving a combination of hematopoietic and viral vector gene transfer therapies were not protected from neurodegneration and axonopathy in both cerebellum and spinal cord. This evidence indicates the requirement of sufficient and widespread GALC expression in CNS and rescue of cerebellum and spinal cord in the therapeutic intervention of murine model of GLD. In this study, we have optimized intracranial delivery of AAV2/5-GALC to the neocortex, hippocampus and cerebellum, instead of the thalamus as was previously conducted, of twitcher mice. The CNS-targeted AAV2/5 gene transfer effectively dispersed GALC transgen along the neuraxis of CNS as far as the lumbar spinal cord, and reduced the accumulation of psychosine in the CNS of twitcher mice. Most importantly, the treated twitcher mice were protected from loss of oligodendrocytes and purkinje cells, axonopathy and marked gliosis, and had significantly improved neuromotor function and prolonged lifespan. Collectively, our study addresses that this severe GLD murine model can be rescued by CNS-targeted AAV5 gene transfer and supports the development of AAV-mediated intervention as a potential therapeutic approach for GLD.
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Tan, Mohd Adi Firdaus. "Metachromatic leukodystrophy improved diagnosis and prognosis / Mohd Adi Firdaus Tan." 2006. http://hdl.handle.net/2440/22369.
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"Date submitted: 28 March 2006"<br>Includes addendum attached to back page and cover.<br>Bibliography: leaves 225-250.<br>xv, 250 leaves : ill. (some col.), tables, plates ; 30 cm.<br>Title page, contents and abstract only. The complete thesis in print form is available from the University Library.<br>Thesis (Ph.D.)--University of Adelaide, School of Paediatrics and Reproductive Health, Discipline of Paediatrics, 2006
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"Characterization of arylsulfatase A from different forms of metachromatic leukodystrophy patients." Tulane University, 1994.
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Arylsulfatase A (ASA) is a lysosomal enzyme whose primary physiological function is desulfation of cerebroside sulfate. Deficiency of ASA leads to metachromatic leukodystrophy (MLD) with different ages of onset. Some healthy individuals also show low ASA activity which is sometimes indistinguishable from that in MLD patients. These so-called ASA pseudodeficient (PD) individuals bring complications to the diagnosis of MLD. This dissertation attempts to establish a model to study the relationships between genotype and phenotype for the PD individuals and MLD patients with different ages of onset Mutations were identified in the ASA-deficient variants using the direct solid-phase sequencing method. These base changes lead to amino acid alterations. Three novel mutations were confirmed by restriction analyses of more than 100 normal alleles. ASA was purified from human liver and the rabbit anti-human ASA antiserum was obtained. Attempts were made to produce monoclonal antibodies against ASA and two of the monoclonal antibodies reacted with ASA in Western blots. ASA from fibroblasts of normals and ASA-deficient variants was partially purified. Western blots were performed after sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), isoelectric focusing (IEF), and dot blot of fractions from the purification procedure. The Western blotting patterns were different among the controls and ASA-deficient variants The K$\sb{\rm m}$ and heat-inactivation profiles were established from normals and an PD individual. The K$\sb{\rm m}$ of the PD individual was three times that of the controls. The half-lives of ASA from normal subjects were three times that of the PD individual at 60$\sp\circ$C and double that of the PD individual at 65$\sp\circ$C The genotype and phenotype relationships were analyzed and this study provides information for diagnosis, prognosis, genetic counseling and gene therapy for MLD patients<br>acase@tulane.edu
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Karumbayaram, Saravanan [Verfasser]. "Biochemical and cell culture approach to study the molecular aspects of metachromatic leukodystrophy / vorgelegt von Saravanan Karumbayaram." 2006. http://d-nb.info/978382951/34.
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"Developing Mesenchymal Stromal Cell Therapy for Neurodegenerative Diseases using the Murine Models of Globoid Cell Leukodystrophy and Multiple Sclerosis." Tulane University, 2015.
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As a novel therapy for neurodegenerative diseases, transplantation of multipotent mesenchymal stromal cells (MSCs) requires extensive optimization in animal models before being implemented in clinical trials. It is a goal of our laboratory to understand the mechanism of action of these cells and to improve their therapeutic efficacy. To address these goals, this study aims to optimize the cell dosage, cell type, administration route and timing, and/or donor age for stem cell therapy in two mouse models of demyelinating diseases: globoid cell leukodystrophy (GLD; Krabbe’s disease) and experimental autoimmune encephalomyelitis (EAE). GLD is a neurodegenerative lysosomal storage disease caused by the deficiency of galactocerebrosidase (GALC). Accumulation of toxic byproducts in myelin producing oligodendrocytes leads to the demyelination of neurons and increase in brain inflammation. The twitcher mouse model of GLD was used to test the therapeutic effects of MSCs after injection through intracerebroventricular (ICV) or intraperitoneal (IP) routes. Weekly MSC IP injections and single IP GALC-transduced MSC injections were performed. Other twitcher mouse cohorts received temporal vein (TV) or intracerebral (IC) injections of GALC-containing adeno-associated virus serotype 9 (AAV9-GALC) with or without IP MSC injections. All GLD affected mice treated with peripheral MSC and/or vector therapy had extended lifespans with improved motor function. The ameliorating effects of MSCs were related to their potent anti-apoptotic and anti-inflammatory effects on the peripheral and central nervous systems. These results indicate a promising future for peripheral administration of MSCs and vectors as non-invasive, adjunct therapies for patients affected with GLD. A similar study was performed using the EAE mouse model of multiple sclerosis (MS), which is a demyelinating disease due to an autoimmune reaction to myelin. The results demonstrated that biological age of the donor reduces the ability of MSCs to alleviate symptoms and improve pathology in the EAE mouse model. Upon transplantation, the young, but not old, MSCs provided neuroprotective effects through immunomodulation and remyelination in the central nervous system (CNS). The age-related therapeutic differences corroborate recent findings that biologic aging occurs in stem cells and highlight the potential need for allogeneic transplantation of MSCs in older MS patients.<br>acase@tulane.edu
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Ramakrishnan, Hariharasubramanian [Verfasser]. "Effects of transgenic overexpression of Polysialytransferase (PST) on myelination and impact of increased sulfatide accumulation in Arylsulfatase A deficient mice and its relevance to Metachromatic leukodystrophy / vorgelegt von Hariharasubramanian Ramakrishnan." 2007. http://d-nb.info/986757845/34.
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Stumpf, Sina Kristin. "Therapeutic approaches for two distinct CNS pathologies." Doctoral thesis, 2018. http://hdl.handle.net/21.11130/00-1735-0000-0005-13E7-1.
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Tétreault, Martine. "Études de nouvelles maladies neurogénétiques chez les Canadiens français." Thèse, 2013. http://hdl.handle.net/1866/10127.
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Depuis déjà plusieurs décennies, nous sommes en mesure d'identifier les mutations responsable de diverses maladies mendéliennes. La découverte des gènes responsables de ces maladies permet non seulement un meilleur diagnostic clinique pour ces familles, mais aussi de mieux comprendre les mécanismes physiopathologiques de ces maladies ainsi que mieux définir la fonction normale des gènes causales. Ultimement, ces découvertes mènent à l'identification de cibles thérapeutiques pour le traitement de ces maladies. Les progrès technologiques sont depuis toujours un facteur très important dans la découverte de ces gènes mutés. De l'approche traditionnelle de clonage positionnel en passant par la première séquence du génome humain et maintenant les technologies de séquençage à grande échelle, de plus en plus de maladies ont maintenant une entité génétique. Dans le cadre de ce projet de doctorat, nous avons utilisé tant les approches traditionnelles (leucodystrophies) que les nouvelles technologies de séquençage (polyneuropathie douloureuse) qui ont mené à l'identification du gène causal pour plusieurs de nos familles. L'efficacité de ces deux approches n'est plus à démontrer, chacune d'entre elles possèdent des avantages et des inconvénients. Dans le cadre de ces projets, nous avons utilisé la population canadienne-française connue pour ces effets fondateurs et la présence, encore aujourd'hui, de grandes familles. Les différents projets ont permis d'établir certains avantages et inconvénients quant à l'utilisation de ces techniques et de la population canadienne-française. Dans le cadre d'un phénotype assez homogène et bien défini comme celui du projet leucodystrophie, l'approche traditionnel par gène candidat nous a permis d'identifier le gène causal, POLR3B, sans trop de difficulté. Par contre, pour les autres projets où nous sommes en présence d'une hétérogénéité clinique et génétique une approche non-biaisée utilisant le séquençage exomique a obtenu un plus grand succès. La présence de grandes familles est un grand avantage dans les deux approches. Dans le projet polyneuropathie douloureuse, une grande famille originaire du Saguenay-Lac-St-Jean nous a permis d'identifier le gène NAGLU comme responsable suite à l'exclusion des autres variants candidats par analyse de ségrégation. Comme NAGLU était déjà associé à un phénotype qui diffère sur plusieurs points à celui de notre famille, une approche traditionnelle n'aurait pas été en mesure d'identifier NAGLU comme le gène causal. Dans l'analyse de nos données de séquençage exomique, nous avons observé que plusieurs variants rares, absents des bases de données, étaient partagés entre les différents individus Canadiens français. Ceci est probablement dû à la démographie génétique particulière observée chez les Canadiens français. En conclusion, les technologies de séquençage à grande échelle sont avantageuses dans l'étude de maladies hétérogènes au niveau clinique et génétique. Ces technologies sont en voie de modifier l'approche d'identification de gènes en permettant une analyse de génétique inversée, c'est-à-dire de la génétique vers la clinique.<br>Since many decades we are able to identify mutations responsible for Mendelian diseases. The identification of the causative gene not only allows a better diagnostic to these families, but also allows a better understanding of the pathophysiological mechanisms of these disorders and of the normal function of a gene. Ultimately, those discoveries lead to the identification of therapeutic targets that will enable clinicians to treat these diseases. Technological progress has forever driven gene identification. Starting with traditional approaches like positional cloning passing through the first sequence of the human genome and finally the recent high-throughput sequencing technologies, more and more diseases are now linked to a genetic cause. During my PhD, I had the opportunity to use traditional approaches (leukodystrophy) as well as new sequencing technologies (painful sensory polyneuropathy) which both led to gene identification for some of our families. The efficiency of these approaches is well known, each of them has advantages and disadvantages. In these projects, we used the french-canadian population well known for its founder effect and the presence, still today, of large families. Working on these projects allowed us to establish advantages and disadvantages concerning the use of those technologies and the french-canadian population. In the presence of a homogenous and well defined phenotype, like the leukodystrophy project, the traditional approach enables us to rapidly identify the causative gene (POLR3B). On the other hand, the unbiased exome sequencing approach has had more success for diseases characterized with clinical and genetic heterogeneity. Large families are a great advantage for both methods. In the painful sensory polyneuropathy project, a large family originated from the Saguenay-Lac-St-Jean region allowed us to identify the NAGLU gene as responsible for the disease after exclusion of the candidate variants by segregation analysis. NAGLU has already been associated with a phenotype that differs in many points with the clinical features observed in our family. In this case a traditional approach would have failed to identify NAGLU as the causative gene. In the analysis of our exome sequencing results, we observed many rare variants absent from databases but shared between french-canadian individuals. This enrichment in rare variants is probably due to the particular genetic demography of Quebec. In conclusion, high-throughput sequencing technologies are advantageous in the study of clinically and genetically heterogeneous diseases. These technologies are changing the gene identification approach towards reverse genetics, meaning genetics towards clinic.